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通过对噬菌体展示的黑素细胞 cDNA 表达文库的血清筛选鉴定出白癜风中的自身抗原。

Autoantigens in vitiligo identified by the serological selection of a phage-displayed melanocyte cDNA expression library.

机构信息

Department of Human Metabolism, School of Medicine, University of Sheffield, Sheffield, UK.

出版信息

J Invest Dermatol. 2010 Jan;130(1):230-40. doi: 10.1038/jid.2009.207.

DOI:10.1038/jid.2009.207
PMID:19587696
Abstract

Vitiligo is an acquired idiopathic hypomelanotic disorder characterized by circumscribed depigmented macules resulting from the loss of cutaneous melanocytes. Although the exact cause of vitiligo remains obscure, autoimmunity may play a role in the development of the disease. The present study was undertaken to investigate the applicability of phage display technology to identify B-cell autoantigens in vitiligo. A melanocyte cDNA phage display library was subjected to rounds of enrichment with vitiligo patient IgG. Subsequently, enriched IgG-binding peptides representing putative autoantigens were identified by sequencing their encoding cDNAs. Radioimmunoassays were used to confirm the immunoreactivity of vitiligo patient (n=61) and control (n=28) sera to several of the putative autoantigens. Non-segmental vitiligo patient sera (n=53) were positive for antibody (Ab) reactivity to gamma-enolase (8%); alpha-enolase (9%); heat-shock protein 90 (13%); osteopontin (4%); ubiquitin-conjugating enzyme (15%); translation-initiation factor 2 (6%); and GTP-binding protein, Rab38 (15%). Ab reactivity to at least one of the previously unknown autoantigens was detected in 51% of patients with non-segmental vitiligo. In contrast, Ab reactivity in a group of patients with segmental vitiligo (n=8) was not demonstrated. Overall, the study indicated that the targets of autoantibodies in vitiligo patients can be revealed by employing the methodology of phage display.

摘要

白癜风是一种获得性特发性色素减退症,其特征是皮肤黑素细胞丧失导致局限性脱色斑。虽然白癜风的确切病因仍不清楚,但自身免疫可能在疾病的发展中起作用。本研究旨在探讨噬菌体展示技术在鉴定白癜风 B 细胞自身抗原中的应用。用白癜风患者 IgG 对黑素细胞 cDNA 噬菌体展示文库进行多轮富集。随后,通过测序其编码 cDNA 来鉴定代表潜在自身抗原的富集 IgG 结合肽。放射免疫测定法用于确认若干潜在自身抗原的白癜风患者(n=61)和对照(n=28)血清的免疫反应性。非节段性白癜风患者血清(n=53)对γ-烯醇化酶(8%)、α-烯醇化酶(9%)、热休克蛋白 90(13%)、骨桥蛋白(4%)、泛素结合酶(15%)、翻译起始因子 2(6%)和 GTP 结合蛋白 Rab38(15%)具有抗体(Ab)反应性。在 51%的非节段性白癜风患者中检测到至少一种先前未知的自身抗原的 Ab 反应性。相比之下,在一组节段性白癜风患者(n=8)中未检测到 Ab 反应性。总的来说,该研究表明,通过采用噬菌体展示技术,可以揭示白癜风患者自身抗体的靶标。

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